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DEVELOPMENT AND VALIDATION OF AN HPTLC METHOD FOR BAKUCHIOL QUANTIFICATION IN PSORALEA CORYLIFOLIA SEEDS USING CENTRAL COMPOSITE DESIGN

MOHD AMIR 1, DEEPAK KHURANNA 2, WASIM AHMAD 3, SHOWKAT R MIR2, MOHAMMAD RAISH 4, SHEIKH MANSOOR5, AJAZ AHMAD 6, MOHD MUJEEB2*

1Department of Natural Products & Alternative Medicine, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia
2School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi-62, India
3Department of Pharmacy, Mohammed Al-Mana College for Medical Sciences, Safaa, Dammam 34222, Saudi Arabia
4Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
5Department of Plant Resources and Environment, Jeju National University, 63234, Republic of Korea
6Department of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia

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Bakuchiol (meroterpenic phenol) is a well-known active constituent present in Psoralea corylifolia Linn (Leguminosae family). The aim for the current study was to determine, develop and validate a new, simple, selective, precise, and accurate highperformance thin layer chromatographic (HPTLC) method for the assessment of bakuchiol in different extracts of P. corylifolia. A pre-coated aluminium plate with silica gel 60 F254 was used for the thin layer chromatographic separation of bakuchiol, eluted with toluene: ethyl acetate: acetic acid (10:1.0:0.1; v/v/v). The plate was subsequently dried, and a Camag TLC scanner III was used to perform densitometric scanning at 262 nm. Selected mobile system gave well defined and sharp chromatogram of bakuchiol at Rf value of 0.48 ± 0.07. Linearity, accuracy, precision, specificity, limit of detection (LOD), and limit of quantification (LOQ) were among the ICH guidelines parameters that were used in the method development and validation. Additionally, a central composite design (CCD) was used in robustness testing. With a fine correlation value (R 2 ) of 0.999, an excellent linear association between concentrations (200 - 2000 ng/band) and areas of the chromatogram was recorded. Intraday and inter-day precision evaluations showed the relative standard deviation 0.68 - 2.77. The percentage of recovery was found to be 98.42% - 101.91%. Results from recovery studies demonstrated outstanding reproducibility and dependability of the method. The LOD and LOQ were determined to be 181.74 ng/spot and 550.74 ng/spot, respectively. The selected factors evaluated in the robustness testing by CCD were found to have a significant effect on the peak area. Among all extracts, the methanolic extract showed maximum concentration of bakuchiol. HPTLC-based identification and quantification of bakuchiol in plant species as well as other plant based and modern medicine formulations may now be done with fresh insights provided by validated standards-based methodologies.